KR20170033295A - Sequencing device - Google Patents

Abstract

A method of making a reagent includes inserting a cartridge into a device. The cartridge includes a plurality of reagent enclosures disposed in the cavity of the cartridge and exposing the ports to the exterior of the cartridge. Each reagent enclosure comprises a reagent vessel containing a reagent, an internal cavity defining a compressible volume, and an opening defined through the reagent vessel as an internal cavity. The method includes connecting a plurality of fluid ports to the openings of a plurality of reagent enclosures; Applying a solution through the fluid ports to at least partially fill the plurality of reagent enclosures; And cycling the pressure of the cavity portion so that, for each of the reagent enclosures, during pressure increase, the solution enters the interior cavity of the reagent vessel, engages the reagent and compresses the compressible volume, The compressible volume is reduced and the reagent is released through the opening.

Description

[0001] SEQUENCING DEVICE [0002]

The present disclosure relates generally to improved sequence analysis devices.

Biomedical research is increasingly dependent on sequencing to improve biological research and medicine. For example, biologists and zoologists rely on sequence analysis to study the origin of animal migration, species evolution, and traits. The medical community depends on sequence analysis to study the origins of the disease, susceptibility to drugs, and the origin of the infection. However, sequence analysis has traditionally been an expensive process and thus its performance has been limited.

By referring to the accompanying drawings, it is believed that the present disclosure may be better understood and many features and advantages of the present disclosure may become apparent to those skilled in the art.
BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a view showing an example of a sequence analyzing apparatus. Fig.
Fig. 2 is a schematic diagram showing an example of a sequence analysis instrument; Fig.
Fig. 3 is a schematic diagram showing an example of a sequence analysis instrument; Fig.
4 is a perspective view illustrating an exemplary reagent storage device;
5 is a perspective view illustrating an example of a container;
6 is a cross-sectional perspective view illustrating an example of a container.
7 is a schematic exploded view illustrating an example of a container.
8 is a detailed perspective view of an example of a container.
9 is a perspective view illustrating an exemplary reagent storage device;
10 is a perspective view illustrating an example of a container;
11 is a cross-sectional perspective view illustrating an example of a container.
Figures 12, 13, 14, 15, and 16 illustrate exemplary cartridges for sealing one or more enclosures.
17 and 18 are schematic diagrams showing an example of a method.
19 is a schematic exploded view of an example of a valve.
20 is a schematic cross-sectional view illustrating an example of a valve.
21 is a schematic sectional view for explaining an example of a valve.
22 is a schematic sectional view for explaining an example of a valve.
23 and Fig. 24 show an example of a manifold and a cartridge. Fig.
25 is a view showing an example of a fluid circuit;
26 is a flow chart of an example of a method of producing a reagent solution;
27 is a flowchart of an example of a method of measuring an analyte;
28 is a flow chart of an example of a method of manufacturing a reagent cartridge.
Using the same reference numerals in different drawings represents the same or similar items.

In one exemplary embodiment, the sequencing system includes a mechanism for carrying out processes for receiving the semiconductor sequence analysis chip and identifying the base sequence. Specifically, the apparatus can accommodate a reagent cartridge, a washing solution, and a semiconductor sequence analyzing chip. The instrument may include a user interface, such as a touch screen user interface, and may include a controller and an arithmetic logic controller that controls acquisition of data from semiconductor sequence analysis that facilitates transfer of the reagents and wash fluid to the semiconductor sequencing chip and identification of the base sequence Circuit.

An exemplary instrument includes a reagent cartridge receptacle, an additional receptacle for receiving a cleaning buffer cartridge, and a chip clamp for receiving a semiconductor sequencing chip. In addition, the device includes a touch screen user interface. Such appliances include: limited sequencer touch points; ease of use (including closed-pump drive systems); no high-quality water utilization; intuitive graphical user interfaces; Operation, fast one-day runtime, integrated onboard operation for primary data analysis, dual mode operation for RUO or diagnostic mode (Dx), small benchtop footprint, scalability Upgradable chassis) or low cost, or any combination thereof.

In particular, the mechanism may include a compressor to provide an internally generated gas pressure. The reagent may be provided in a preloaded cartridge to limit user interaction with reagent production. Similarly, the cleaning liquid may be provided as a plug-and-play cleaning liquid in the cartridge or the like. For example, the pH of the wash liquor may be stabilized using a solid buffer.

The flow rate of the reagents in the system can be controlled, for example, using dynamic flow control using a pinch valve regulator. The system may also include a wash liquor for performing post-run washing. The apparatus may include an internal server for processing data received from the semiconductor sequencing chip. Alternatively or additionally, the system may provide its output data for connection to an external server to process the output data. In addition, the internal computing system is configurable and upgradeable.

The system can utilize the pressure driven liquid flow using an internal gas supply without using an external gas supply. In particular, the reagent cartridge system may utilize a liquid or lyophilized nucleotide in a separate enclosure or bag. The enclosure can deflate any initial air content by pressurizing the chamber outside the bag and flowing (flowing) air from the bag to the waste. The cleaning liquid can be applied in the bag. Residual air bubbles can rise to the top and be purged with waste. Mixing is achieved by pressurizing and rapidly depressurizing the chamber outside the bag to pressurize and depressurize the liquid in the bag. The liquid or lyophilized nucleotides are contained in a mixer (reagent vessel) that releases a solution containing the nucleotides in response to the reduced pressure to cause mixing of the nucleotides in the bag.

As the external gas pressure is applied to the back enclosure, the internal scrubbing liquid flows into the mixer and the internal compressible volume in the mixer is compressed. When the gas pressure outside the bag is quickly released, the sealing pressure in the compressible volume causes the cleaning liquid to move outwardly through the nozzles to the outside, thereby mixing the liquid containing the nucleotides in the bag.

Dynamic flow control can be achieved by utilizing a pinch flow regulator (pinch valve regulator) when applying a flow through a flow cell of a semiconductor sequencing chip. This dynamic flow control reduces the use of resistance tubing coils and reduces potential clogging. The flow rate can be programmed and adjusted by adjusting the control pressure in the pinch flow regulator.

The system may utilize a solid buffer system, such as a ceramic buffer system, for example, particulate titanium dioxide. The wash liquor reagent may be provided in a disposable bottle that is easily mixed. The wash liquor may contain a solid buffer to remove the automatic pH routines and provide long term pH stability. The particulate titanium dioxide can be easily put into a bottle by a filter, so that the fine particles can be restricted in the system. Also, the primary cleaning bottle can utilize packaging that is low permeability to gas and thus low permeability to CO ₂ acidification. The vessel may also have a CO ₂ absorption packet to further limit exposure to carbon dioxide.

Semiconductor sequencing chips can be housed in a chip clamp that mounts the chip in the system and connects the chip to the fluid system. The chip clamp may include an integrated squeegee valve manifold and remove the tubing connection. The chip clamp may also include a reference electrode, an integrated chip temperature control, and an integrated manifold heating.

The mechanism may also depend on the reagent cartridge, so that contact and handling by the user may be limited. Simple cartridges are mounted and contain five containers or enclosures (for nucleotide and bead retrieval or pH adjusting reagents). The cartridge may also include an enclosure port for rapidly depressurizing / depressurizing the cartridge utilizing, for example, an exhaust valve on a manifold connected to the reagent cartridge. Optionally, a silencer is attached to the exhaust valve to reduce noise associated with rapid depressurization of the cartridge.

The apparatus further includes an expandable and configurable electronic device. In particular, it is possible to exchange computation power and memory. The system may further include a support for the RFID tag on, for example, a semiconductor sequencing chip or a cartridge.

Exemplary reagent cartridges may contain different nucleotide solutions and color-coding ports for bead detection or pH adjusting solutions. The reagent cartridge may also include input and output ports for the CO ₂ scrubber. The cartridge may also include an RFID tag that identifies, for example, the lot number and expiration date.

The cartridge may include individualized chambers into which reagents are applied or a CO ₂ scrubber is inserted. The reagent containers are inserted into the enclosure or reagent pouch secured to the lid in the cartridge through the cartridge lid.

In order to produce an exemplary reagent cartridge, a fitting including, for example, a low density polyethylene / polyethylene terephthalate film is affixed within the pouch. The pouches are then attached to the lid to assemble the cartridge base, the lid, the port gasket, and other components, followed by attaching the lid to the base, and also inserting the gasket into the lid. The reagent vessel includes a mixer body into which a foam member is inserted. Attach the mixer cap to the mixer body. In one example, the mixer cap may comprise a lyophilized reagent or a liquid reagent. In another example, the reagent can be frozen or stored in a porous ceramic or polymer film. The assembled reagent mixers and scrubbers can then be attached to the reagent cartridge assembly. The RFID tag can be attached to the cartridge, and the cartridge can be stored in a box for shipment.

The reagent may be applied to the mixer or cartridge in lyophilized form or in the form of a frozen liquid. For example, lyophilized nucleotide pellets may be formed and then inserted into mixers. As another example, nucleotides can be dried on filter paper that is integrated with bug mixers. In another example, the nucleotides can be dried directly on a compressible foam or in a cap (second portion) of a mixer (reagent vessel).

In one example, Figure 1 illustrates an exemplary instrument 100 for sequencing. The instrument 100 may include a connector for receiving a container of the buffer solution 104, a clamp 106 for receiving the sequencing device, and a manifold for receiving the reagent cartridge 102. The sequencing instrument 100 also includes a fluid flow, data retrieval from the sequencing device, an arithmetic circuit that controls data interpretation, and a user interface 108. In a specific example, the sequencing device is a pH or a sensitive ion device including, for example, a plurality of ion sensitive field effect transistors (ISFETs).

As a specific example, the sequencing instrument includes circuitry for controlling fluid flow in the system. In the example illustrated in FIG. 2, the mechanism 200 includes connections to the cartridge 202. The reagent may flow from the cartridge 202 through the fluid circuit 204 to the sequencing device 206. The fluid passing through the fluid circuit 204 may be selectively directed to the waste container 212 or may be directed through the sequencing device 206 to the waste container 210 via the pinch flow regulators 208 or 230 have. In an alternative embodiment, one waste container may replace the waste containers 210 or 212. One exemplary embodiment of the cartridge 202 is shown in Figures 12-16. One exemplary embodiment of a pinch flow regulator is shown in Figures 19-22. Exemplary fluid circuits are shown in Figures 3 and 25.

As further described below, a solution in the container 226, such as a buffer solution, can flow through the valve 228 and can be used to produce a reagent solution in the reagent enclosure 214. [ The reagent solution 214 may selectively flow to the fluid circuit 204 and to the sequencing device 206 or the waste containers 210 or 212. The solution in the solution reservoir 226 may selectively flow through the valve 232 to the fluid circuit 204 to selectively clean the fluid circuit 204 and optionally the reagent sequencing device 206 from the reagent solution And can function as a washing liquid for washing. The buffer solution in the solution container 226 may be selectively pumped in the system. Alternatively, the buffer may be actuated by pressure and may be supplied, for example, using air through inlet 234.

The system 200 may include a compressor 216 that compresses the gas or air to flow through the scrubber cartridge 220 optionally contained in the reagent cartridge 202. For example, the scrubber cartridge 220 may include components such as soda lime to remove carbon dioxide from the air. The pressurized air can be stored and supplied to the system using the re-centric 218. For example, pressurized air can be used to pressurize the solution vessel 226. [ In another example, pressurized air may be supplied to the cavity of cartridge 202 to press reagent enclosures 214 and selectively drive reagent solutions into fluid circuit 204 through valves 222 . In a further example, compressed air from the receptacle 218 may be used to clean the fluid circuit 204 through the valve 224. The pressurized air causes the remaining liquid in the fluid circuit 204 to enter the reagent enclosures 212 through the waste container 212, the sequencing device 206 into the deoxygenated waste container 210, 214).

Figure 3 shows one more detailed embodiment of the fluid circuit. Figure 3 illustrates a system employing an enclosure 614, which is, for example, a reagent reservoir to perform pH-based nucleic acid sequence analysis. Each electronic sensor in the device generates an output signal. The fluid circuit allows a plurality of reagents to be delivered to the reaction chambers.

3, the system is connected to reagent reservoirs 614 and is connected to a waste reservoir 620 and is connected to a fluid reservoir 620 that connects the fluid node 630 to the inlet 638 of the biosensor 634 for fluid communication. And a fluid circuit 602 connected to the biosensor 634 by a passage 632. [ The reagent solution produced and mixed from the reservoirs 614 can be moved to the fluid circuit 602 by various methods including pressure, pumps such as a syringe pump, gravity feed, etc., and by controlling the valves 650 Is selected. The reagents from the fluid circuit 602 can move to the waste containers 620, 636. The control system 618 may include controllers for valves 650 that generate signals for opening and closing through the electrical connection 616.

The control system 618 also includes controllers for other components of the system, such as the wash fluid valve 624 and the reference electrode 628, which are connected to the control system by the electrical contact 622. [ The control system 618 may also include control and data acquisition functions for the biosensor 634. In one mode of operation, the fluid circuit 602 is configured such that between the selected reagent flows, the fluid circuit 602 is prime, cleaned by the wash fluid 626 and biosensor 634 is washed with the wash fluid 626, Under the programmed control of the control system 618, the sequence of the selected reagents (1, 2, 3, 4, or 5) is transmitted to the biosensor 634. The fluid entering the biosensor 634 flows out through the outlet 640 and is deposited in the waste container 636. For example, a similar setup may be used for an optical sequencing system with a photodiode or CCD camera.

As a specific example, wash fluid 626 may be a buffered suspension comprising solid buffer microparticles. The buffer suspension may be filtered using the filter 660 before entering the fluid circuit 602 or the sensor 634. As a further example, a buffered suspension may be applied to the reagent reservoirs 614 through a filter 662 to form reagent solutions from the reagent concentrate in the reagent reservoirs. Alternatively, the filters 660 and 662 may be the same filter. For example, the reagent concentrate is a liquid concentrate. In another example, the reagent concentrate is a dried concentrate such as a lyophilized reagent (e.g., a lyophilized nucleotide). Alternatively, the illustrated filters 660 and 662 may be combined. In another example, the filters may be located downstream of reagent reservoirs 614, such as between reagent reservoirs 614 and valves 650.

4 is a diagram illustrating an exemplary reagent storage device 1100. FIG. In one example, the reagent storage device 100 may include an enclosure 1110. The container 1120 is disposed within the enclosure 1110. In one example, the enclosure 1110 may be a flexible enclosure. A flexible enclosure, such as a sealable flexible back enclosure, can be pressurized and depressurized by externally applied pressure, for example, by applying a pressurized gas to the exterior surface of the flexible enclosure. Alternatively, the enclosure may be rigid so that externally applied gas pressure is not readily converted to fluid pressure within the enclosure 1110.

The reagent storage device 1100 may also include a fluid port 1130 connected to a fitting 1160 attached to the enclosure 1110 to provide fluid access to the interior of the enclosure 1110. The fluid port 1130 may be connected to the fitting 1160 to seal the enclosure 1110 from the external environment after the container 1120 is inserted. The enclosure 1110 may be heat sealed to itself and the fitting 1160, for example, and otherwise sealed by the fluid port 1130.

The container 1120 may include one or more arms 1140 and a flange 1150. The arms 1140 can place the container 1120 in the enclosure 1110, e.g., approximately at the center, to evenly distribute the reagent within the enclosure 1100. [ The flange 1150 can be provided for convenient assembly of the container 1120. In one example, the arm 1140 is flexible. For example, the arm 1140 may be formed of wire or polymer material. Alternatively, the arm 1140 may be rigid. Alternatively, the arm 1140 and the flange 1150 are not limited to those shown in FIG. 4 and may include a structure for positioning the container within a predetermined position or orientation within the enclosure 1110. The container 1120 may be connected directly or indirectly to the fluid port 1130 or alternatively may be located at a suitable distance away from the fluid port 1130 as shown in FIG. The enclosed enclosure 1100, including the enclosure 1110 and the container 1120, provides for simplified storage and transfer of reagents in the container 1120.

5 is a perspective view illustrating an example of the container 1200. FIG. The container 1200 may include a first portion 1210 and a second portion 1220 coupled to the first portion 1210. Elements such as optional compressible members and reagents may be inserted into the container 1200 before the first portion 1210 is connected to the second portion 1220. [ In one example, the second portion 1220 may be a cap that slides over or otherwise covers a portion of the first portion 1210 to form an inner cavity. In another example, the second portion 1220 may be an insert that slides into the first portion 1210. The second portion 1220 may be formed by screwing or otherwise screwing the second portion 1220 onto the first portion 1210, any suitable attachment mechanism including a locking mechanism, adhesive, or any other suitable attachment mechanism And may be connected to the first portion 1210 by an attachment mechanism.

In one example, the inner cavity defines a compressible volume. The compressible volume is compressed in response to fluid pressure and does not dissipate or leave the internal cavity of the container 1200. The compressible volume may comprise a compressible gas volume or may be a compressible member such as an elastic polymer or foam.

The container 1200 may define a passage 1230 that provides fluid communication between the interior cavity of the container 1200 and the exterior of the container 1200. In one example, one or more passageways 1230 can be defined up to the inner cavity. These passages 1230 may be drilled through a first portion or a second portion of the vessel 1200. Alternatively, the second portion 1220 may include a passageway 1230, or may extend beyond the region where the second portion 1220 engages the first portion 1210 to form a passageway 1230 Lt; / RTI > slots.

The one or more arms 1240 can be connected to the first portion 1210 to position the container 1200 as desired within the enclosure. The flange 1250 may be connected to the second portion 1220 to assist in attaching the second portion 1220 to the first portion 1210 or to place the container 1200 away from the lowermost portion of the enclosure.

6 is a cross-sectional perspective view illustrating an example of the container 1300. As shown in Fig. The vessel 1300 defines an internal cavity 1320 and defines a passageway 1330 that provides fluid communication between the internal cavity 1320 and the exterior of the vessel 1300. The passageway 1330 may be drilled through the vessel 1300. In another example, the cap or insert may include a slot that extends beyond the region of engagement with the container 1300 and forms the passageway 1330. [ The container 1300 includes a first portion 1310 and a first portion 1310 that allow elements such as compressible member 1340 and reagents to be inserted into the interior cavity 1320 of the container 1300 And a second portion 1350 coupled thereto.

Internal cavity 1320 defines a compressible volume. The compressible volume is the volume that is compressed in response to the pressure to match the pressure and can expand in response to the reduced pressure to provide a counter force against the fluid pressure. In one example, the compressible volume is compressed to match the pressure of the fluid entering the inner volume without extinguishing the inner cavity or leaving the inner cavity, and pressurizing the fluid from the inner cavity 1320 in response to the reduced pressure of the fluid And a compressible gas. Optionally, the compressible volume may comprise a compressible member 1340. The compressible member 1340 can be compressed upon pressurization and returns to its previous form substantially upon decompression. For example, the compressible member 1340 can be a foam material. In particular, the compressible member 1340 can be a closed cell foam of elastomeric material. In one example, the compressible member may comprise a polyurethane foam.

In one example, the reagent may be disposed within the second portion 1350. The reagent may be a lyophilized nucleotide or the like. In another example, the reagent is a solution that is absorbed in a porous metal, ceramic, or polymer sponge-like material or frit. Optionally, the reagent solution can be frozen. In an alternative, the reagent may comprise a pH adjusting reagent such as an acid or base.

The one or more arms 1360 can be connected to the first portion 1310 to position the container 1300 as desired within the enclosure. The flange 1370 or other suitable attachment may be attached to the second portion 1350 to assist in attaching the second portion 1350 to the first portion 1310 or to place the container 1300 in the enclosure.

7 is a schematic exploded view explaining an example of the container 1400. Fig. The container 1400 can include a first portion 1410 and a second portion 1420 (e.g., an insert) coupled to the first portion 1410 to facilitate the movement of the selectively compressible member 1430, So that the elements can be inserted into the container 1400. The second portion 1420 may be secured to the first portion 1410 by sliding it into the first portion 1410 or by screwing it with the first portion 1410. One or more flexible arms 1440 may be connected to the first portion 1410 to position the container 1400 in the enclosure. Flange 1450 may be coupled to second portion 1420 to assist in coupling engagement of second portion 1420 and first portion 1410 or to position container 1400 in the enclosure.

In one example, the second portion 1420 may define one or more slots 1460. The first portion 1410 and the second portion 1420 can be coupled to leave a portion of one or more of the slots 1460 exposed so that the gap between the interior cavity of the container 1400 and the exterior of the container 1400 One or more passageways may be provided.

8 is a detailed perspective view for explaining an example of the container 1500. Fig. A detailed perspective view of the container 1500 may define a passage 1510 that provides fluid communication between the interior cavity of the container 1500 and the exterior of the container 1500. The end of the container 1500 includes a fitting 1530 for receiving the insert 1540. The insert 1540 includes a hole or slot that is not covered when the insert 1540 is attached to the fitting to form the passageway 1510. Alternatively, the insert 1540 can include an aperture, notch, mesh, pore, or any other suitable feature to provide fluid communication to the fitting 1530. Flange 1520 can be connected to container 1500 to control insertion portion 1540 when attached to fitting 1530 and to position the container within the enclosure.

9 is a perspective view illustrating an exemplary reagent storage device 1600. FIG. The reagent storage device 1600 includes an enclosure 1610. The container 1620 is disposed within the enclosure 1610. Enclosure 1610 may be a flexible enclosure as discussed above. For example, a flexible enclosure may be a sealable flexible back enclosure that can be pressurized and depressurized externally through fluid or gas pressure. Alternatively, the enclosure 1610 may be a rigid enclosure. Enclosure 1610 may be sealably coupled to a sealing structure 1670, such as a fitting having a bore 1680, such as a central bore. The container 1620 can be connected to an arm 1640 that can be connected to the fluid port 1630 and can be inserted through the bore 1680 of the fitting 1670.

Fluid port 1630 provides fluid access to the interior of enclosure 1610 through bore 1680. The fluid port 1630 may be connected to the fitting 1670 of the reagent storage device 1600 or the sealing structure from the external environment after the container 1620 is inserted. The arm 1640 connects the vessel 1620 to the fluid port 1630 to position the vessel 1620 approximately at the center within the enclosure 1610, for example, to uniformly distribute the reagent within the enclosure 1610 . The arm 1640, the container 1620, and the fluid port 1630 can be one integrated piece. In one example, the fluid flows selectively through the bore 1680 of the fitting 1670 through the fluid port 1630 to the enclosure 1610 along the arm 1640. The arm 1640 can position the container 1620 with or without a flange 1650.

10 is a perspective view for explaining an example of the container 1700. Fig. The container 1700 can include a first portion 1710 and a second portion 1720 connected to the first portion 1710 so that elements such as a selectively compressible member can be inserted into the container 1700 do. Fluid port 1730 is connected to container 1700 and provides fluid access to the enclosure into which container 1700 is inserted. The arm 1740 may connect the first portion 1710 and the fluid port 1730 to position the container 1700 in the enclosure.

In one example, the second portion 1720 is an insert for engaging the first portion 1710. In another example, the second portion 1720 forms a cap to cover the end of the first portion 1710. Fluid may flow through the aperture 1770 of the port 1730 and into the aperture 1760 along the arm 1740. The fluid port may include a gasket to facilitate sealing. The flange 1750 may be connected to the second portion 1720.

11 is a cross-sectional perspective view illustrating an example of the container 1800. Fig. The container 1800 defines an internal cavity 1820 and defines a passageway 1830 that provides fluid communication between the interior cavity 1820 and the exterior of the container 1800. The container 1800 can include a first portion 1810 and a second portion 1850 connected to the first portion 1810 to define a compressible volume. In one example, the compressible member 1840 and elements such as reagents may be inserted into the interior cavity 1820 of the container 1800.

In one example, the second portion 1850 is a cap for attaching over the end of the first portion 1810. In another example, the second portion 1850 is an insert for attaching to the fitting of the first portion 1810. In one example, passage 1830 is formed in second portion 1850, for example, as a hole or slot.

The reagent may be disposed within the second portion 1850. The reagent may be a lyophilized nucleotide or the like. In another example, the reagent can be a porous metal, a ceramic, or a nucleotide solution that is absorbed by a polymer sponge or frit. In a further example, the reagent may be frozen. In a further example, the reagent may comprise a pH adjusting reagent such as an acid or base.

Fluid port 1860 is connected to container 1800 and provides fluid access to the enclosure into which container 1800 is inserted. For example, fluid entering opening 1890 may pass through passages 1895 and into the enclosure. The arm 1840 may be connected to the first portion 1810 and the fluid port 1860. The flange 1880 can also be connected to the second portion 1850 to position the container 1800 in the enclosure.

The reagent storage device may be inserted into a cartridge or case having a cavity. In one example, the pressure can be varied within the cavity to change the pressure of the liquid in the flexible enclosure and thus affect the pressure in the container. Alternatively, pressure may be applied to the interior of the enclosure through opening 1890. In one example, one or more of the enclosures may be integrated into the case. The case may define one or more pressure chambers that can be pressurized and released from the enclosures.

12, the cartridge or case 1900 includes a lid 1902 and a body 1904. [ The lid 1902 can receive fluid ports 1906, 1908, 1910, 1912, or 1914 of the containers that are inserted into the flexible enclosures. The container may contain different reagents. For example, each container may contain nucleotides, or may comprise a pH adjusting reagent. The lid 1902 may also include a port 1916 for providing or releasing pressurized gas to control the pressure externally of each of the enclosures and thereby control the pressure in the enclosure have. The walls of the base 1904 and lid 1902 can be configured to pressurize the cavity in the cartridge 1900, for example, with pressurized gas or air. In one example, the cartridge 1900 may be labeled with a bar code or radio frequency identification (RFID) tag.

As shown in Figures 12 and 13, lid 1902 may include access ports 1918, 1920 for applying gas or air through the scrubber cartridge. In particular, the system can utilize external air, so that external air can be applied through port 1918 and clean gas or air can be received through port 1920. In particular, the scrubber cartridge may contain absorbent materials for capturing carbon dioxide or water. Carbon dioxide can be removed from the air to prevent acidification of the liquid components when carbon dioxide is diffused into the enclosures or when air is used in other parts of the system.

In a further example, lid 1902 may also include alignment features 1924 or 1926. With such alignment features, access to the ports 1906, 1908, 1910, 1912, 1914, 1916, 1918, or 1920 can be aligned with the manifold to limit damage to the manifold, It is possible to provide a proper coupling between the case 1900.

14, 15 and 16, the body 1904 can define individual cavities 2126 in which each enclosure 2128 is disposed and in which the nucleotide container 2130 is inserted. In one example, each enclosure 2128 is disposed in separate cavities 2126, and each container 2130 is attached through a lid 1902 to engage lid 1902 at the fluid port of container 2130. Fittings 2134 of enclosures 2128 can engage lid 1902.

The lid 1902 may define a headspace that provides communication between the pressurized gas inlet port 1916 and each of the cavities 2126. [ Alternatively, the cavity may be an open cavity free of individualized cavities 2126 and may provide a single cavity to which a pressurized gas may be applied to apply pressure to the enclosures 2128. [ As shown in FIG. 15, the body 1904 may include a chamber 2232 for receiving a scrubber cartridge, for example, to remove carbon dioxide from the air.

As seen from the top view, body 1904 includes individualized cavities 2126 as shown in Fig. In addition, the body may include a sealing structure 2340 to separate the scrubber cartridge input and output from the rest of the pressure of the body 1904. Further, the inner sealing portion 2342 can be utilized to separate the input pressure of the air entering the scrubber cartridge from the output pressure of the air deviating from the scrubber cartridge. The body 1904 may also include a sealing structure 2344 that engages an opposing sealing structure on the lid 1902 to provide a separate interior space that includes cavities that can be pressurized or depressurized.

The containers may contain nucleotide reagents or other reagents. In particular, individual containers within a cartridge system may include one of four nucleotides. The system may also include a container within the enclosure containing pH adjusting reagents. As a specific example, the cartridge optionally comprises a pH adjustment reagent vessel and containers and enclosures that integrate each of the four nucleotides (A, G, C, or T). For example, the reagents are in a dried form. For example, lyophilized nucleotides can be stored in a container. In another example, the reagent solution may be absorbed within a porous metal, ceramic, or polymer sponge-like material or frit. In a further example, the reagent solution may be frozen in a container or in a porous sponge-like material in which the reagent solution is absorbed.

The enclosures described herein can be applied to make reagent solutions. Assembly of the enclosure includes inserting the container into the enclosure and sealing the container within the enclosure to the fluid port. The one or more enclosures may also be secured within the volume of the case, and the case includes a gas port for providing external gas pressure to the fixed enclosures. The enclosures can be inserted just prior to mixing, which provides flexibility in the case as a final assembly step or in the selection of reagents.

Alternatively, the enclosure may be secured to the lid prior to inserting the containers containing the reagent. Reagent containers can be inserted through the lid and the fluid ports of the containers can engage the lid. The lid can be secured to the base after securing the enclosures to the lid or after inserting the containers through the lid into the enclosures.

Pressurization and depressurization of the fluid in the enclosures is controlled by increasing and decreasing the gas pressure of the volume of the case through the gas port.

The method of making a reagent solution includes filling an enclosure, such as any of the enclosures described herein, including a container and a reagent, with a predetermined amount of fluid through the fluid port of the enclosure. The fluid in the enclosure is then pressurized to flow into the interior cavity of the container or through the passageway of the container. The fluid can be directly pressurized through the port. In another example, the fluid may be pressurized by applying an external pressure to the enclosure using, for example, gas or other fluid pressures. Pressurizes the member or compressible volume in the interior cavity of the container while the fluid fills a portion of the volume of the interior cavity.

For example, the fluid flows into the inner cavity of the container and compresses the compressible volume or member until the pressure in the inner cavity applied to the compressible volume or member is approximately equal to the pressure in the enclosure external to the container .

The fluid in the enclosure is depressurized after reaching a predetermined pressure. The compressible volume or member is decompressed to expand and release the fluid and reagent from the inner cavity into the enclosure external to the container. The mixture of reagent and fluid exiting the passageway creates eddy currents and turbulence in the back enclosure that is sufficient to mix the reagent with the fluid. Upon depressurization, the pressure in the inner cavity as applied by the compressible volume or member is greater than the pressure in the enclosure outside the container. The fluid and the reagent are released from the inner cavity through the passageway until the pressure in the inner cavity is approximately equal to the pressure in the enclosure outside the container to provide a well mixed reagent solution.

Pressurization can be performed by increasing the gas pressure outside the flexible enclosure. In one embodiment, the enclosure may be disposed within the case. The pressure in the enclosure can be controlled by increasing / decreasing the gas pressure in the case outside the enclosure. Proper mixing of reagent and fluid can be achieved through repeated cycles of pressurization and depressurization. After mixing is complete, the fluid and the reagent are released through the fluid port of the enclosure.

17 and 18 illustrate an exemplary method of assembling a reagent cartridge. For example, the fitting 702 may be secured to the back enclosure to form the enclosure 708. A plurality of enclosures 708 may be connected to the cartridge lid 706 as illustrated in 714 and may be inserted into the cartridge base 704 when the cartridge lid 706 is secured to the cartridge base 704. [ . The port gasket 710 may be secured to the cartridge lid to enable the compressed air system to be connected to the cartridge. In a further example, the port gasket 712 may be secured to the cartridge cover 706 to allow access to the CO ₂ scrubber.

Referring to FIG. 18, reagent vessels may be formed by inserting a selectively compressible member into a first portion 716 of the reagent vessel, as indicated by reference numeral 722. The second portion 718 may be secured to the first portion 716 to form the reagent vessel 728. Optionally, the reagent is applied to the second portion 718. Alternatively, the reagent is inserted into the first portion at 722.

The scrubber vessel 720 may be filled with a scrubbing reagent to remove CO ₂ , as illustrated at 726, for example. A plurality of reagent vessels 728 and a scrubber vessel 720 may be inserted into the reagent cartridge through the lid as illustrated by reference numeral 730. [ The ends of the reagent cartridge containers 728 are fed into the interior of the reagent enclosures through the cartridge lid. The scrubber vessel 720 is inserted into a separate compartment of the cartridge which allows air to flow in and out through the lid without affecting the pressure of the remaining cavities of the cartridge. The fluid port gasket 724 is secured onto the reagent vessel and optionally secured onto the scrubber vessel to provide fluid tight access to the reagent vessel or scrubber vessel when secured within the manifold and instrument.

Figure 19 provides a schematic exploded view of an example of a pinch valve regulator 3100. [ Valve 3100 includes a housing base 3110 and a housing cover 3120 disposed over the housing base 3110. The diaphragm 3130 is disposed between the housing base 3110 and the housing cover 3120. A pinch plate 3140 is disposed between the diaphragm 3130 and the base 3110. In operation, the pinch plate 3140 is configured to limit the flow of fluid through the fin tube 3150 (as shown more clearly in Figures 20, 21 and 22) And moves with respect to the housing base 3110. One or more gaskets 3160 and 3170 can be disposed between the housing base 3110 and the housing cover 3120 to prevent fluid leakage and ensure smooth valve operation.

Figure 20 provides a schematic cross-sectional view of an example of a pinch valve regulator. The valve 3200 includes a housing 3210 and a housing cover 3220 disposed over the housing 3210. Housing base 3210 includes a pinch structure 3214 that protrudes into lower cavity 3212 and lower cavity 3212. The housing base 3210 includes a gas inlet 3230 that provides external access to the lower cavity 3212. The base fluid inlet 3232 provides an external access path connected to one end of the pinch tube 3240 in the lower cavity portion 3212. The other end of the pinch tube 3240 is connected to the base fluid outlet 3234. Accordingly, the pinch tube 3240 provides fluid communication between the base fluid inlet 3232 and the base fluid outlet 3234. The pinch tube 3240 extends between the pinch point 3252 of the pinch plate 3250 and the pinch structure 3214.

In one example, the pinch structure 3214 includes a rectangular prism that extends into the lower cavity 3212. As shown, the rectangular prism has a round top. In another example, the rectangular prism may have a flat top. Alternatively, the prism may have a point-like structure such as a triangular prism. In general, the pinch structure 3214 forms a counter structure in which the pinch point 3252 fixes and punches the pinch tube 3240. [

The base fluid outlet 3234 is then connected to the cover fluid inlet 3236 and communicates with the cover fluid inlet 3236 between the upper and lower cavities to provide a fluid path through the housing base 3210 and the housing cover 3220. [ Respectively. Cover fluid inlet 3236 is in fluid communication with cover fluid outlet 3238 through fluid path 3270. The cover fluid outlet 3238 provides external access to the fluid path 3270 from the housing cover 3220. The diaphragm 3260 is positioned between the housing base 3210 and the housing cover 3220 to fluidly isolate the lower cavity 3212 from the upper cavity 3222 defined between the cover 3220 and the diaphragm 3260. [ .

The housing cover 3220 defines an upper cavity 3222 in which the fluid path 3270 is disposed. Alternatively, the gasket 3280 may define a portion of the lower cavity 3212 or a portion of the upper cavity 3222. The pinch plate 3250 can be disposed within the cavity region defined by the gasket 3280 or the housing cover 3220. [ Base fluid outlet 3234 and cover fluid inlet 3236 are in fluid communication with gasket 3280 and diaphragm 3260. Alternatively, the base fluid outlet 3234 and the cover fluid inlet 3236 may be fluidly connected outside the housing base 3210 or the housing cover 3220. The diaphragm 3260 provides separation between the lower cavity 3212 and the upper cavity 3222. The pinch plate 3250 is disposed within the housing covers 3220 and the cavities 3212 and 3222 defined in the housing base 3210. The pinch plate 3250 includes a pinch point 3252 disposed on the opposite side of the pinch structure 3214. Pinch point 3252 is shown as having a rounded tip. Alternatively, the pinch point 3252 may have a sharp tip. The pinch plate 3250 moves relative to the housing base 3210 to limit fluid flow through the pinch tube 3240 based on the fluid pressure in the fluid path 3270 and the gas pressure in the lower cavity 3212, The tube 3240 is picked up.

The valves described herein operate to adjust fluid flow as a function of gas pressure in the lower cavity. Fig. 20 shows the valve structure before applying the fluid to the valve 3200, and Figs. 21 and 22 show the equilibrium state of the valve through which the fluid flows through the valve 3300 at a flow rate based on the input gas pressure. An embodiment of an operating pinch valve will be described later with reference to Figs. 20, 21 and 22. Fig.

Gas pressure is applied to the gas inlets 3230 and 3330 of the valves to pressurize the lower cavities 3212 and 3312 with an input / reference gas pressure. The pressurized lower cavity portion applies an upward force to the pinch plates 3250 and 3350 and the diaphragms 3260 and 3360 toward the housing covers 3220 and 3320. Fluid is applied to base fluid inlet 3332 and flows through pinch tube 3340, base fluid outlet 3334, cover fluid inlet 3336, fluid path 3370, cover fluid outlet 3338, Flows sequentially. The fluid flowing through the housing cover 3320 applies a downward force to the diaphragm 3360 and the pinch plate 3350 toward the housing base 3310. The diaphragm 3360 moves toward the housing base 3310 and applies a downward force to the pinch plate 3350 as the fluid pressure in the fluid path 3370 increases with respect to the gas pressure in the lower cavity portion 3312 . In particular, diaphragm 3360 causes pinch point 3352 to point toward pinch structure 3314 in response to a difference between the fluid pressure of upper cavity 3322 and the gas pressure of lower cavity 3312. For example, the diaphragm 360 may include a pinch structure (not shown) in response to an increase in fluid pressure in the upper cavity 3322 relative to the gas pressure in the lower cavity 3312 to limit fluid flow in the pinch tube 3340 Point 3352 to cause the pinch point 3352 to point to the pinch point < RTI ID = 0.0 > 3314. < / RTI &

As the pinch plate 3350 moves toward the housing base 3310, the pinch point 3352 causes the input gas pressure to oppose the fluid pressure in the upper cavity 3322 and thus the constant fluid from the valve 3300 A downward force is applied to the pinch tube 3340 until the flow velocity is provided to restrict the flow of the fluid to the tube 3340 against the pinch structure 3314 or to restrict the flow of the fluid through the punch tube 3340 and the upper cavity 3322 Causing a descent. Figure 22 shows valve 3300 in which directional arrow 3380 represents the fluid flow path through valve 3300. [

The pinch maneuvering force of the diaphragm drive pinch valve is such that the output fluid pressure is adjusted by the input gas pressure. By setting the pressure of the lower gas cavity 3312 to a known value, pressure and fluid flow out of the housing cover 320 are controlled. In this way, the valve self-adjusts to reach an equilibrium state and can provide a desired constant fluid flow. In summary, the output fluid pressure at the cover fluid outlet follows the input gas pressure at the gas inlet, and can be independent of the fluid pressure at the base fluid inlet.

Figures 23 and 24 illustrate an exemplary sequencing system that includes a manifold 806 to receive a cartridge 804. The manifold may be driven up and down using an actuator 808. [ Alternatively, the waste bin 802 may be located within the apparatus and fluidly connected to the cartridge 804, for example, as schematically shown in FIG.

As shown in FIG. 24, a plurality of fluid ports 810 can be connected to the tubing and fluid circuit, such as the fluid portion, schematically shown in FIG. The cartridge 804 may optionally include an RFID tag readable by a mechanism. Connecting the manifold 806 to the cartridge 804 can be prevented or allowed based on reading of the RFID tag.

Figure 25 illustrates another embodiment of a fluid circuit of the present invention that accommodates five input reagents in a planar circuit structure. 25 is a top view of a transparent body or housing 4300 including a fluid circuit 4302. Fig. The housing may be constructed of a variety of materials including metals, glass, ceramics, plastics, and the like. The transparent material includes polycarbonate, polymethylmethacrylate and the like. The inlets (or input ports) are connected by passages to respective connector slots (e.g., 4370) located on the lowermost side of the housing where the reagents enter the fluid circuit 4302. The inlets are in fluid communication with the passages (e.g., 4353), and these passages are again connected to the curved passages. Each curved pathway consists of two legs identified for the curved pathway at the "T" junction 4356. One leg is the inner leg connecting each of its inlets to the node (or the multipurpose central port 4301), and the other one is the outer leg connecting each of its inlets to the waste passage (or ring 4340). As discussed above, the cross-sectional area and length of the inner and outer legs of the curved passageways may be selected to achieve the desired balance of flow at the "T" junctions at node 4301. [ Through the passage, the waste passage (or channel) 4340 is in fluid communication with a waste port 4345 connected to a waste reservoir (not shown) by a connector slot on the lowermost side of the body. The node 4301 is in fluid communication with the port 4363 by a passageway 4361, shown in phantom, external to the body 4300 in this embodiment. In other embodiments, passageway 4361 may be formed in the body such that connector slots for node 4301 and port 4363 are not required. The port is connected by a passage 4363 to a cleaning fluid inlet where a "T" junction is formed and is connected to the connector slot, thereby providing a conduit for the flow cell, reaction chamber, Figure 25 illustrates a mode of using a fluid circuit to distribute fluids to a flow cell. The operating modes are implemented by valves 4350 associated with each of the input reagents and the wash fluid. In the first mode of operation (the selected reagent valve is open, all the other reagent valves are closed and the scrubber valve is closed), the selected reagent is delivered to the flow cell, the second mode of operation (All of the reagent valves are closed and the flush valve is open), the fluid circuit is primed to deliver the selected reagent, the third mode of operation (all reagent valves are closed, the flush valve is open) ), All passages of the fluid circuit are cleaned. As discussed above, what is associated with each inlet port can be opened so that fluid can enter fluid circuit 4302 through each of its respective inlets (as shown for valve 4352) 4352) that can be closed to prevent fluid from entering circuit 4302 (as shown for all but the valves 4352, 4352). In each case, as the valve at the inlet opens and the remaining valves (including the scrubber valve) are closed, as shown for the inlet 4370 in Figure 25, the fluid is forced through the passage 4354 to the "T" And the fluid is divided into two streams at this junction and one stream is directed to the waste passage 4340 and then to the waste port 4345 and the remaining stream is directed to the node 4301. From node 4301, this second flow is again divided into a number of flows, one of which leaves node 4301 through path 4361, then to path 4363, to the flow cell, Flows into each of the passages connecting the inlet port 4301 to the remaining inlets, and then flows into the waste passage 4340 and the waste port 4345. The latter flow carries any material that diffuses or leaks from the remaining inlets and passes through the rest of the inlets that direct it to the waste port 4345. The sequence of different reagents may be directed to the flow cell by opening the valves of the selected reagents and simultaneously closing all of the non-selective reagents and the wash fluid valves. In one embodiment, such a sequence may be implemented by a sequence of operating modes of the fluid circuit, such as cleaning, reagent xl priming, reagent xl transfer, wash, reagent x2 priming, reagent x2 transfer, In the reagent priming mode, all reagent inlet valves except the valve corresponding to the selected reagent, such as the reagent delivery mode, are closed. However, unlike the reagent delivery mode, the wash fluid valve is opened and the relative pressure of the selected reagent flow and wash fluid flow is selected such that the wash fluid flows through the passageway 4361 to the node 4301, And escape through all the passages leading to the waste passage 4340 except for the passage leading to the reagent inlet.

As shown in FIG. 26, a method 900 for manufacturing a reagent solution in a cartridge includes inserting a cartridge into the apparatus, as indicated by reference numeral 902. For example, the cartridge may be inserted under the manifold or under the mechanism shown in Fig. 1, similar to the embodiments shown in Fig. 23 or Fig.

Optionally, the system may check the cartridge alignment features as indicated by reference numeral 904. For example, the cartridge may include a structure that indicates proper positioning of the cartridge within the apparatus. As further indicated by reference numeral 906, the cartridge may optionally include a Radio Frequency Identification Tag (RFID) that can be read by a mechanism. Based on the test of the cartridge alignment features or the reading of the RFID tag of the cartridge, the mechanism may selectively engage the cartridge using a manifold.

For example, the mechanism may connect a plurality of fluid ports secured to the manifold to the cartridge, thereby providing a fluid connection with the remainder of the mechanism, as indicated at 908. The mechanism may also connect a gas system, such as a compressed air system, to the cartridge, as indicated by reference numeral 910. In one example, a connection to the gas system is incorporated into the manifold connecting the fluid ports to the reagent cartridges. In particular, the system can be connected to scrubber inlets and outlets and selectively connected to cavities defining the space between reagent enclosures.

As indicated by reference numeral 912, the cartridge cavity may be pressurized. The enclosure or reagent enclosure can be evacuated by pressing the cartridge cavity. Following the venting of the enclosure, the cartridge cavity can also be depressurized.

A solution (e. G., A buffer) may be applied through the fluid ports as indicated by reference numeral 914. In particular, solutions can be applied universally to each of the reagent enclosures. Following application of the solution, the pressure in the cartridge cavity can be cycled. Such cycling may, for example, utilize the mechanisms described above with respect to Figures 4-16 to cause mixing of reagents in the enclosures.

Figure 27 shows an exemplary method 9204 for cleaning the system after measuring the analyte. For example, as indicated by reference numeral 922, pressure may be applied to the cartridge cavity to pressurize the reagent enclosures.

As indicated by reference numeral 924, the reagents can selectively flow from the reagent enclosures by opening valves connecting the individual reagent enclosures to the fluid circuit. In particular, the reagent may flow sequentially from selected individual reagent enclosures that are selectively separated by a flow of wash liquid from separate containers.

The pressure in the cartridge cavity can provide a driving force for the reagents from the reagent enclosures and the individual reagents can be selected by selectively opening the valve associated with the reagent enclosure while the flow rate, Can be controlled on the downstream side of the fluid circuit. In particular, the flow can be controlled on the downstream side of the fluid circuit, for example, by utilizing the pinch flow regulator to control the flow to the waste container.

The system can then be cleaned by applying pressurized air through the scrubber of the cartridge and flowing the pressurized, scrubbed air through the fluid circuit 930, as indicated by reference numeral 928. [ The pressurized air can drive the fluid from the fluid circuit, for example, through the sensor device and the pinch flow regulator, to the waste containers. In a further example, the pressurized air may drive the reagent fluids back through the respective associated valves into the reagent enclosures in the cartridge. In this case, the cartridge cavity can be depressurized, allowing the pressurized air to drive the reagent fluids back into the reagent enclosures.

As shown in FIG. 28, a method 940 of manufacturing a reagent cartridge includes attaching a plurality of reagent enclosures to a cartridge lid, as indicated at 942. Provides a schematic exemplary method that is illustrated or illustrated in greater detail in Figs. 17 and 18. Fig. The cartridge lid can be secured to the cartridge base, as indicated by reference numeral 944, and a plurality of reagent containers can be inserted into the plurality of enclosures through the cartridge lid, as indicated by reference numeral 946 .

The system or instrument may be integrated into the process flow for sequencing. For example, the system may be utilized in conjunction with the One Touch 2 (registered trademark) or Ion Chef (registered trademark) system for performing template manufacturing. Following the template preparation, the instrument can be utilized for sequencing. The framework can be configured to perform initial analysis, or outsource initial analysis and interpretation to the cloud or external server.

The mechanism may be configurable. For example, the system may include one or more central processing units, a configurable amount of RAM, an upgradeable graphics processing unit, and an exchangeable storage device including 1 to 12 TB. The apparatus is configured to accept different sequencing chips and perform sequencing using these chips. The system may also be upgraded to access external servers for analysis and interpretation of data received from the sequencing chip.

The multiple chips supported by the instrument can also support multiple assays, allowing for a different number of readings, read lengths, base output and application. As such, the system is versatile and useful in a variety of research areas.

Such a system is provided for the preferred sequencing runs, including output from a high-precision run comprising a P1 proton chip containing 19.6 Gb or 15.4 Gb.

In a first aspect, a method of making a reagent includes inserting a cartridge into a device. The cartridge includes a plurality of reagent enclosures disposed within the cavity of the cartridge and exposing the ports to the exterior of the cartridge. Each reagent enclosure includes an opening defined by an interior cavity through the reagent vessel and a reagent vessel containing an internal cavity and reagent defining a compressible volume. The method includes connecting a plurality of fluid ports to the openings of a plurality of reagent enclosures; Applying a solution through the fluid ports to at least partially fill the plurality of reagent enclosures; And cycling the pressure of the cavity so that during each of the reagent enclosures the solution enters the internal cavity of the reagent vessel, is combined with the reagent, compresses the compressible volume, During pressure reduction, the compressible volume is reduced and the reagent is released through the opening.

In one example of the first aspect, the method further comprises pressing the cavity to remove gas from the plurality of enclosures prior to applying the solution.

In another embodiment of the above examples and the first aspect, the reagent vessel further comprises a compressible member disposed in a compressible volume.

In the above examples and additional examples of the first embodiment, the reagent comprises a nucleotide.

In the above examples and a further example of the first aspect, the method further comprises sensing the position of the cartridge prior to connecting the plurality of fluid ports.

In another embodiment of the above examples and the first aspect, the method further comprises reading the identification tag of the cartridge by a mechanism. For example, the method may further comprise connecting a plurality of fluid ports based on the reading.

In the above examples and a further example of the first aspect, the cartridge further comprises a scrubber, and the method further comprises connecting the gas system to the scrubber. For example, the scrubber is a CO ₂ scrubber. In another example, cycling the pressure includes applying gas into the cavity through the scrubber.

In a second aspect, a method of detecting an analyte by a device comprises applying pressure to a cartridge connected to the device. The cartridge includes a plurality of reagent enclosures disposed within the cavity of the cartridge and exposing the ports to the exterior of the cartridge. Each reagent enclosure includes an opening defined by an interior cavity through the reagent vessel and a reagent vessel containing an internal cavity and reagent defining a compressible volume. The method includes selectively flowing a reagent from a reagent enclosure of a plurality of reagent cartridges from the cartridge to a waste container via a fluid circuit, a sensor, and a pinch flow regulator; And controlling flow to the waste container using a pinch flow controller.

In one example of the second aspect, applying pressure to the cartridge includes flowing gas into the cavity of the cartridge through the scrubber of the cartridge.

In another example of the above examples and the second aspect, the sensor comprises a sensitive field effect transistor.

In a further example of the above examples and the second aspect, applying pressure comprises compressing air with a compressor of the apparatus.

In a third aspect, a method of cleaning a device includes applying pressurized air through a scrubber of the cartridge; And flowing the scrubbed pressurized air through the fluid circuit, wherein a portion of the scrubbed pressurized air pressurizes the fluid through the fluid circuit towards the reagent enclosures of the cartridge, Pressurize to waste container through regulator.

In a fourth aspect, a method of manufacturing a reagent cartridge includes attaching a plurality of enclosures to a cartridge lid; Securing the cartridge lid to the cartridge base, wherein the cartridge lid defines a plurality of openings, the opening of the cartridge lid providing access to the interior of any of the plurality of enclosures; And inserting a plurality of reagent vessels into the plurality of enclosures, wherein any one of the plurality of reagent vessels extends through the opening into the interior of the enclosure, and the reagent vessel includes a reagent, an interior defining a compressible volume, A cavity portion, and an access portion defined as an internal cavity portion through the reagent container.

In one example of the fourth aspect, the reagent comprises a nucleotide.

In another example of the above examples and fourth aspect, the nucleotide is disposed on the porous material in the inner cavity of the reagent vessel.

In a further example of the above examples and fourth aspect, the method further comprises inserting the scrubber into the cartridge prior to securing the cartridge cover.

In a fifth aspect, a system includes a cartridge manifold connected to a cartridge and providing fluid communication between a plurality of reagent enclosures and a fluid circuit, the cartridge manifold providing fluid communication between a compressed gas system, a cartridge cavity, The fluid circuit comprising: a sequencing device in fluid communication with the fluid circuit; A pinch flow regulator in fluid communication with the sequencing device; And a waste container in fluid communication with the pinch flow regulator.

It is noted that not all of the above described activities of the coarse description or examples are required, that some of the specific activities may not be necessary, and that one or more additional activities may be performed in addition to the activities described above. Also, the order in which activities are listed is not necessarily the order in which they are performed.

In the foregoing specification, the concepts have been described with reference to specific embodiments. However, it will be understood by those of ordinary skill in the art that various modifications and changes may be made without departing from the scope of the present invention as set forth in the following claims. Accordingly, the specification and figures are to be regarded in an illustrative rather than a restrictive sense, and all such modifications are intended to be included within the scope of the present invention.

As used herein, the terms "comprise", "comprises", "includes", "including", "has", " having "," having ", or any other variation thereof, are intended to encompass non-exclusive inclusions. For example, a process, method, article, or apparatus that includes a list of features is not necessarily limited to this feature, and may include other features not expressly stated or inherent to such process, method, article, or apparatus . In addition, unless the context clearly dictates otherwise, "or" refers to "inclusive-or" and not "exclusive-or." For example, a condition A or B is when A is true (or is present) and B is false (or not present); A is false (or not present) and B is true (or is present); And where both A and B are true (or present).

Also, the use of the singular representation is taken to illustrate the components and elements described herein. This is done for convenience only and provides a general sense of the scope of the invention. This description should be construed to include one or at least one, and the singular forms also include the plural unless the context clearly indicates otherwise.

Benefits, other advantages, and solutions to problems have been described with regard to specific embodiments. However, it is to be understood that any feature (s) that may generate or clarify advantages, advantages, solutions to problems, and any benefit, advantage, or solution, Should not be construed as being a part, or an essential feature.

It will be appreciated by those of ordinary skill in the art, after reading the specification, that certain features have been described herein in the context of separate embodiments for clarity and may also be provided in conjunction with one embodiment. Conversely, various features described in the context of an embodiment may be provided individually or in any sub-combination for brevity. Also, the values referred to in the reference ranges include all individual values within the range.

Claims (20)

As a method for producing a reagent,
Inserting a cartridge into a device, wherein the cartridge includes a plurality of reagent enclosures disposed in a cavity of the cartridge and exposing a port to the exterior of the cartridge, each reagent enclosure being compressible A reagent vessel including an inner cavity defining a volume and a reagent, and an opening defined through the reagent vessel to the inner cavity;
Connecting a plurality of fluid ports to the openings of the plurality of reagent enclosures;
Applying a solution through the fluid ports to at least partially fill the plurality of reagent enclosures; And
Cycling the pressure of the cavity such that for each of the reagent enclosures during cycling the solution enters the internal cavity of the reagent vessel and engages the reagent and the compressible volume And compressing, during pressure reduction, the compressible volume is reduced and the reagent is released through the opening. 2. The method of claim 1, further comprising pressurizing the cavity to remove gas from the plurality of enclosures prior to applying the solution. 3. The method of claim 1 or 2, wherein the reagent vessel further comprises a compressible member disposed within the compressible volume. 4. The method according to any one of claims 1 to 3, wherein the reagent comprises a nucleotide. 5. The method of any one of claims 1 to 4, further comprising sensing the position of the cartridge prior to connecting the plurality of fluid ports. 6. The method of any one of claims 1 to 5, further comprising reading the identification tag of the cartridge by the mechanism. 7. The method of claim 6, further comprising connecting the plurality of fluid ports based on the reading. 8. The method of any one of claims 1 to 7, wherein the cartridge further comprises a scrubber, the method further comprising connecting a gas system to the scrubber. The method of claim 8, wherein the scrubber is a CO ₂ scrubber. 9. The method of claim 8, wherein cycling the pressure comprises applying gas into the cavity through the scrubber. A method for detecting an analyte by a mechanism,
Applying pressure to a cartridge connected to the device, the cartridge comprising a plurality of reagent enclosures disposed within a cavity of the cartridge and exposing a port to the exterior of the cartridge, each reagent enclosure defining a compressible volume A reagent vessel containing an internal cavity and a reagent, the opening defined by the reagent vessel into the internal cavity;
Selectively flowing a reagent from a reagent enclosure of the plurality of reagent cartridges from the cartridge to a waste container via a fluid circuit, a sensor, and a pinch flow regulator; And
And controlling flow to the waste container using a pinch flow controller. 12. The method of claim 11, wherein applying pressure to the cartridge comprises flowing gas through the scrubber of the cartridge into the cavity of the cartridge. 13. The method of claim 11 or 12, wherein the sensor comprises an ion sensitive field effect transistor. 14. A method according to any one of claims 11 to 13, wherein applying pressure includes compressing air with a compressor of the instrument. CLAIMS 1. A method of cleaning an instrument,
Applying pressurized air through the scrubber of the cartridge; And
Flowing the scrubbed pressurized air through a fluid circuit, wherein a portion of the scrubbed pressurized air pressurizes the fluid through the fluid circuit towards the reagent enclosures of the cartridge, And pushing through the pinch flow regulator to the waste container. A method of manufacturing a reagent cartridge,
Attaching a plurality of enclosures to the cartridge cover;
Securing the cartridge lid to a cartridge base, wherein the cartridge lid defines a plurality of openings, the opening of the cartridge lid providing access to the interior of any of the plurality of enclosures; ; And
Inserting a plurality of reagent containers into the plurality of enclosures, wherein any one of the plurality of reagent containers extends through the opening into the interior of the enclosure, the reagent containers defining a reagent and a compressible volume Wherein the reagent container comprises an inner cavity, and wherein the access is defined through the reagent container to the inner cavity. 17. The method of claim 16, wherein the reagent comprises a nucleotide. 18. The method of claim 16 or 17, wherein the nucleotide is disposed on the porous material in the interior cavity of the reagent vessel. 19. The method of any one of claims 16 to 18, further comprising inserting a scrubber into the cartridge prior to securing the cartridge lid. As a system,
A cartridge manifold connected to the cartridge and providing fluid communication between the plurality of reagent enclosures and the fluid circuit, the cartridge manifold providing fluid communication between the compressed gas system, the cartridge cavity, and the scrubber of the cartridge,
The fluid circuit comprising:
A sequencing device in fluid communication with the fluid circuit;
A pinch flow regulator in fluid communication with the sequencing device; And
And a waste container in fluid communication with the pinch flow regulator.

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